PROJECT ABSTRACT This project proposes to evaluate the role of interleukin-10 (IL10) in intrauterine growth restriction (IUGR) on endothelial dysfunction, leading to placental insufficiency in the short-term and adult cardiovascular disease (CVD) in the long-term. IUGR and placental insufficiency impose significant health care burden, and remain poorly understood and untreatable. The mechanisms underlying development and maintenance of fetal placental vascular structures are unknown, and are disrupted in IUGR. IL10, an anti-inflammatory cytokine, protects against endothelial dysfunction in several human and animal applications. The innovation of this project is to understand immunologic mechanisms of fetal placental vascular maintenance via IL10 in IUGR pregnancies. The hypothesis is that dysregulated IL10 signaling pathways and downstream effectors affect vascular maintenance of the placenta, and in IUGR, this dysregulation leads to reprogramming of fetal endothelial cells (ECs), and risk of adult CVD in offspring. The following specific aims are proposed: 1. Determine the effect of IUGR on IL10 signaling mechanisms and downstream effects on cellular maintenance and survival of human fetal endothelial cells isolated from placenta 2. Investigate the in-vivo role of IL10 in placental vascular insufficiency and programming of endothelial dysfunction, as mediated through cellular autophagy and ER stress, in a murine model of chronic IUGR 3. Demonstrate the mechanistic effects of altered IL10 on development of vascular structures, vascular function, autophagy and ER stress in cultured human endothelial cells in-vitro These specific aims will be completed by using several techniques, which include magnetic bead isolation and flow cytometry, immunophenotyping, RNA and protein expression assays, genomic methylation platforms (aim 1), use of animal models (aim 2), and cell culture and lentiviral overexpression/knock-down constructs (aim 3). The expectation is that these studies will show that IL10 receptor pathways are activated in fetal endothelial cells of placenta, and maintain homeostasis of cell survival processes. Low levels of IL10 will increase cellular autophagy and ER stress, leading to endothelial dysfunction. Rescue with IL10 will restore endothelial function and placental vascularity. IUGR will lead to epigenetic modification of vasoregulatory pathways in fetal ECs. Completion of this project will be guided by a team of mentors and collaborators with relevant expertise in fields and techniques key to the proposal. The candidate will undertake formal coursework to address gaps in knowledge. A timeline to evaluate periodic assessment of the candidate's progress is included in this proposal. This project has significant relevance to public health as identification of pathways mediating endothelial function can be targeted for therapeutic intervention to protect the fetus from adverse outcomes.